production of cellulases and hemicellulases by penicillium

ORIGINAL ARTICLE

Production of cellulases and hemicellulases by Penicilliumechinulatum grown on pretreated sugar cane bagasse andwheat bran in solid-state fermentationM. Camassola and A.J.P. Dillon

Institute of Biotechnology, University of Caxias do Sul, Caxias do Sul-RS, Brazil

Introduction

Solid-state fermentation (SSF) is a process whereby an

insoluble substrate is fermented with sufficient moisture,

but without free water (Chahal 1985; Lonsane et al.

1992). This system presents many advantages over sub-

merged fermentation (SmF), including high volumetric

productivity, relatively higher concentration of the prod-

ucts, less effluent generation, requirement for simple

fermentation equipment, etc. (Pandey et al. 1999). Fur-

ther, the ability of SSF to minimize catabolic repression

already has been described for several enzymes (Aguilar

and Huitron 1986; Ramesh and Lonsane 1990, 1991;

Solis-Pereyra et al. 1996; Archana and Satyanarayana

1997; Siqueira et al. 1997; Nandakumar et al. 1999).

In recent years the interest in cellulases and hemicellu-

lases has increased because of many potential applications

for these enzymes. Cellulases and hemicellulases can be

used, for example, in the formulation of washing pow-

ders, in the textile industry (Cavaco-Paulo 1998), in the

pulp and paper industry (Ferreira Filho 1998; Dhillon

et al. 2000), in processes including supplementation of

animal feeds (Mandebvu et al. 1999), extraction of fruit

and vegetable juices, starch processing (Rolle 1998;

Gusakov et al. 2000; Park and Park 2001). Still, the grow-

ing concerns about the potential consequences of a

worldwide shortage of fossil fuels, the emission of green

house gases and air pollution by incomplete combustion

of fossil fuel has also resulted in an increased focus on

the production of bioethanol from lignocellulosics

Keywords

bioethanol, cellulases, hemicellulases,

Penicillium echinulatum, sugar cane bagasse.

Correspondence

Aldo J.P. Dillon, Institute of Biotechnology,

University of Caxias do Sul, Francisco Getulio

Vargas Street 1130, Caxias do Sul-RS, 95070-

560 Brazil. E-mail: [email protected]

2007 0338: received 3 March 2007, revised 2May 2007 and accepted 2 May 2007

doi:10.1111/j.1365-2672.2007.03458.x

Abstract

Aim: To evaluate the solid-state fermentation (SSF) production of cellulase and

hemicellulases (xylanases), by Penicillium echinulatum 9A02S1, in experiments

carried out with different concentrations of the pretreated sugar cane bagasse

(PSCB) and wheat bran (WB).

Methods and Results: This study reports the production of xylanolytic and cel-

lulolytic enzymes by P. echinulatum 9A02S1 using a cheap medium containing

PSCB and WB under SSF. The highest amounts of filter paper activity (FPA)

could be measured on mixtures of PSCB and WB (3289 190 U gdm)1). Thehighest b-glucosidase activity was 5895 258 U gdm)1 on the fourth day.The highest activity for endoglucanases was 28236 123 U gdm)1 on thefourth day, and for xylanases the activity was around 10 U gdm)1 from the

second to the fourth day.

Conclusions: The present work has established the potential of P. echinulatum

for FPA, endoglucanase, b-glucosidase and xylanase productions in SSF, indica-ting that WB may be partially substituted by PSCB.

Significance and Impact of the Study: The incorporation of cheap sources,

such as sugar cane bagasse, into media for the production of lignocellulose

enzymes should help decrease the production costs of enzymatic complexes

that can hydrolyse lignocellulose residues for the formation of fermented

syrups, thus contributing to the economic production of bioethanol.

Journal of Applied Microbiology ISSN 1364-5072

2196 Journal compilation 2007 The Society for Applied Microbiology, Journal of Applied Microbiology 103 (2007) 21962204 2007 The Authors

(Sheehan and Himmel 1999; Zaldivar et al. 2001), and

especially the possibility of using cellulases and hemicellu-

lases to perform enzymatic hydrolysis of the lignocellulo-

sic material (Himmel et al. 1999; Sun and Cheng 2002).

However, in bioethanol production, it is necessary to

reduce the costs of the enzymes used to hydrolyse the raw

material and to increase their efficiency in order to render

the process economically feasible (Sheehan and Himmel

1999). In addition, there is a general interest in obtaining

new, more specific, stable enzymes (Jrgensen et al. 2003)

and to use a cheap source of inducer, such as sugar cane

bagasse, and recycle all or part of the enzymes.

Sugar cane bagasse is an abundant, low-cost lign-

ocellulosic material (Takahashi et al. 2000; Ferrara et al.

2002). The appropriate use of sugar cane bagasse enhan-

ces the value of this material and provides a solution for

the removal of this abundant waste, solving a problem of

the sugar industry and increasing the economic yield

of the process (Gamez et al. 2006). However, this material

needs to be pretreated to break the lignin seal and disrupt

the crystalline structure of cellulose to make cellulose

more accessible to the enzymes that convert the carbohy-

drate polymers into fermentable sugars.

In this context, the aim of the present work was to

evaluate the SSF production of cellulase [filter paper

activity (FPA), endoglucosidase and b-glucosidase] andhemicellulases (xylanases), by Penicillium echinulatum

9A02S1, in experiments carried out with different ratios

of PSCB and WB.

Materials and methods

Micro-organism

The cellulolytic mutant P. echinulatum strain 9A02S1

(DSM 18942) was used in this study. This strain was

obtained by exposing wild type P. echinulatum strain

2HH to ultraviolet (UV) light and hydrogen peroxide

(H2O2) (Dillon et al. 2006). These strains are stored in

the culture collection of the Division of Enzyme and Bio-

mass, Institute of Biotechnology, Caxias do Sul, Rio

Grande do Sul, Brazil. The strain was grown on C-agar

slants (Dillon et al. 2006) for up to 7 days at 28C untilconidia formed, and then stored at 4C until use.

Delignified bagasse

Sugar cane bagasse was thoroughly milled to 110 mm

particle size. The delignification of milled bagasse was car-

ried out using three parts of solution 16% of sodium

hydroxide + 03% hydrogen peroxide + 002% antraqui-none (AQ) for one part of sugar cane bagasse (w w), at120C for 20 min. After autoclaving, the pretreated

bagasse was washed with tap water, next with distilled

water until neutrality, and then dried at 60C.

Enzyme production

Pretreated sugar cane bagasse and WB were used as the

support and main carbon sources. The culture media

consisted of mixtures of different ratios of PSCB and WB,

as mentioned in the results. The controls were performed

with WB and PSCB.

Fermentations were performed in flasks with a

12 3-cm concave base; the flasks were closed with agauze-covered cotton wool plug containing 2 g of dry

mass of production media and 2 ml basal salt solution

containing (in g l)1) KH2PO4, 20; (NH4)2SO4, 13;

CO(NH2)2, 3; MgSO47H2O, 3; CaCl2, 3; FeSO47H2O,0050; MnSO4H2O, 00156; ZnSO47H2O, 0014; andCoCl2, 00020. The flasks were autoclaved at 120C for20 min. Each flask was then inoculated with sufficient

conidial suspension to give a final concentration of

1 106 conidia per gram of dry mass of productionmedia. The medium moisture was adjusted to 67% by

the addition of distilled water. The flasks were incuba-

ted at 28C and 90% humidity for 5 days. Experimentswere carried out with three replicates for each medium

composition and for each incubation time. To extract

the enzymes after incubation, the contents of each flask

were separately added to a 125-ml Erlenmeyer flask

containing 10 ml of distilled water and the pH was

measured and 17 ml of 005 mol l)1 citrate buffer (pH48) was added, mixed, incubated under agitation for30 min at 4C and filtered. The filtrate was assayed forenzymes as described below.

Enzyme assay

The enzymatic activity was analysed on filter paper

(FPA), according to Ghose (1987). The b-glucosidaseactivity was dosed using salicin as the substrate, accord-

ing to Chahal (1985). Endoglucanase activity was deter-

mined according to Ghose (1987) using 2% (w v)carboxymethylcellulose solution in citrate buffer. The

reducing sugar was estimated as glucose equivalent by

the dinitrosalicylic acid (DNS) method according to

Miller (1959). The xylanase activity was measured by

the method of Bailey et al. (1992) using oat spelt xylan.

Reducing sugar was measured by the DNS method

using xylose as standard.

Enzyme units

One unit (U) of enzyme activity was defined as the

amount of enzyme required to release 1 lmol of reducing

M. Camassola and A.J.P. Dillon Production of cellulases and hemicellulases by P. echinulatum

2007 The AuthorsJournal compilation 2007 The Society for Applied Microbiology, Journal of Applied Microbiology 103 (2007) 21962204 2197

sugar from the appropriate substrates per minute under

assay conditions. The enzymatic activities are expressed as

units per gram of dry medium (U gdm)1).

Mycelial mass determination

The quantity of N-acetylglucosamine was determined by

the method described in Reissig et al. (1955) and the

quantity of mycelial mass estimated according to Bitten-

court et al. (2002).

Statistical tests

The results were statistically analysed using analysis of

variance with the Tukey post-test for a P < 005 using thePrismGraphPad program (Graph Pad, San Diego, CA).

Results

Enzyme production

Tests were carried out employing mixtures of different

proportions of PSCB and WB to verify the secretion of

cellulases and xylanases by P. echinulatum strain 9A02S1.

We also performed fermentation experiments with two

control media, one of them containing only WB and the

second containing only PSCB. Wheat bran was used,

because it is a nutrient-rich substrate presenting ideal

conditions to produce cellulases and xylanases. The results

of the enzymatic analysis are expressed as units per gram

of dry medium (U gdm)1) in Fig. 1a,b.

Filter paper activity

Figure 1a shows the results of FPA. It is found that the

control treatment formulated only with PSCB

(10PSCB : 0WB) presented the lowest enzymatic activities

of FPA throughout the experiment, except on the fourth

and fifth days; that the control formulated with WB

(0PSCB : 10WB) presented similar activities. On the

second day, the enzymatic activity of treatment (2PSCB :

8WB) was superior to that of the control (0PSCB :

10WB); although two other treatments (6PSCB : 4WB

and 4PSCB : 6WB) did not show a statistical difference,

they had means higher than this control, while the

treatments formulated with greater amounts of PSCB

a

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(a) (b)

(d)(c)

10PSCB:0WB 8PSCB:2WB 6PSCB:4WB4PSCB:6WB 2PSCB:8WB 0PSCB:10WB

Figure 1 Variation of the filter paper activity (a), b-glucosidases (b), endoglucanases (c) and xylanases (d) in media formulated with different pro-

portions of solid-state pretreated cane bagasse and wheat bran, using strain 9A02S1 of Penicillium echinulatum. PSCB: pretreated sugar cane

bagasse, WB: wheat bran. The numbers shown in the legends indicate the proportion of each medium component used. Values (averages) with

the same letters for the same day do not differ significantly by the Tukey test (P > 0005).

Production of cellulases and hemicellulases by P. echinulatum M. Camassola and A.J.P. Dillon


presented the smallest amount of activity. The highest

enzymatic activity of FPA for the 2PSCB : 8WB

(3289 190 U gdm)1) treatment was obtained on thethird day. However, the 4PSCB : 6WB treatment presen-

ted similar activity and, furthermore, kept up its enzyma-

tic activity until the end of this experiment. In this

sampling, the higher productivities of this experiment

were also determined. Values of (1096 063 U gdm)1

per day) were obtained for treatments 2PSCB : 8WB and

(1040 201 U gdm)1 per day) for 4PSCB : 6WB, whilethe highest productivity obtained by the control treat-

ment (0PSCB : 10WB) was 534 014 U gdm)1per day,on the second day of culture. On the third day also, it

was found that culture 6PSCB : 4WB also presented enzy-

matic titers statistically higher than the control

0PSCB : 10WB, while the culture 8PSCB : 2WB presented

activities similar to this control. On the fourth and fifth

days, all cultures formulated with mixtures of PSCB and

WB presented higher activity than the controls.

b-Glucosidases

The data obtained in the b-glucosidase dosages are plot-ted in Fig. 1b. It was found that the control treatment

10PSCB : 0WB presented the lowest enzymatic activities

of b-glucosidase throughout the experiment. On thesecond day, two cultures (2PSCB : 8WB and

4PSCB : 6WB) presented higher means, one with similar

values (6PSCB : WB) and the other two treatments

(10PSCB : 0WB and 8PSCB : 2WB) with lower activity

than the control treatment 0PSCB : 10WB. During the

sampling performed on the third day of culture, it was

found that treatment 2PSCB : 8WB presented an activity

of (4887 1117 U gdm)1), which enabled higher pro-ductivity of the experiment (1629 339 U gdm)1 perday), followed by treatment 4PSCB : 6WB which presen-

ted a b-glucosidase activity of 4333 690 U gdm)1 andproductivity of 1444 230 U gdm)1 per day. In thissampling, control 0PSCB : 10WB presented enzymatic tit-

ers of 2693 683 U gdm)1, while the other cultures pre-sented less activity. On the fourth day, once again,

treatment 2PSCB : 8WB was outstanding, and enzymatic

titers of 5895 258 U gdm)1 were obtained, followedby treatments 4PSCB : 6WB (4623 1141 U gdm)1)and 6PSCB : 4WB (4439 363 U gdm)1). On the fifthday, all cultures with PSCB had a drop in enzymatic

activity, except for the control treatment 0PSCB : 10WB

which presented increased activity.

In treatments formulated with mixtures of WB and

PSCB, except for treatment 6PSCB : 4WB on the fifth

day of culture, higher enzymatic activity was obtained

in the treatments supplemented with higher proportions

of WB. It should also be pointed out that enzymatic

activities were higher than the control containing only

WB.

Endoglucanases

In solid-state culture, endoglucanase activity was highly

favoured, using WB and PSCB. Activities higher than

200 U gdm)1 were obtained on the third and fourth days

by cultures 6PSCB : 4WB and 4PSCB : 6WB, respectively,

as seen in the data shown in Fig. 1c.

As in the case of FPA and b-glucosidases, the controlculture 10PSCB : 0WB also had the lowest enzymatic

activities of endoglucanase throughout the experiment.

On the second day, cultures 2PSCB : 8WB and

6PSCB : 4WB presented endoglucanase activities that

were statistically superior to the control culture

0PSCB : 10WB, while the enzymatic dosages of culture

4PSCB : 6WB were similar. On the third day, two cul-

tures (4PSCB : 6WB and 2PSCB : 8WB) presented more

endoglucanase activities than control 0PSCB : 10WB. On

the fourth day, only culture 10PSCB : 0WB, formulated

with PSCB exclusively, had less activity than control

0PSCB : 10WB. Similar behaviour was observed on the

fifth day, although with less activity.

The treatment 6PSCB : 4WB behaved strangely during

this experiment. On the third day, there was a sudden

drop in enzymatic activity, followed by a large increment

on the fourth day and a new drop on the fifth day.

Xylanases

Figure 1d shows the enzymatic titers obtained for xyla-

nases. As for the other enzymes analysed, treatment

10PSCB : 0WB presented the lowest enzymatic activities

of xylanases throughout the experiment. On the second

day of the culture, three treatments (6PSCB : 4WB;

4PSCB : 6WB and 2PSCB : 8WB; 919 050 U gdm)1,981 048 U gdm)1 and 995 153 U gdm)1, respect-ively) presented xylanasic activities similar to the control

0PSCB : 10WB (967 019 U gdm)1). On the third day,cultures 4PSCB : 6WB and 2PSCB : 8WB maintained

higher activities than the control 0PSCB : 10WB, while

on the fourth day, the other cultures that presented more

activity than control 0PSCB : 10WB repeated the beha-

viour. Already on the fifth day, the last three treatments

mentioned presented enzymatic activity for xylanases sim-

ilar to the control containing only WB.

As to the pH (Fig. 2a) of different treatments, it was

found that the higher the amounts of WB used, the lower

were the pH values. However, there was no direct rela-

tionship between pH and the enzymatic activities. From

the second day onwards, it was seen that the cultures with

higher proportions of WB (0PSCB : 10WB, 2PSCB : 8WB



and 4PSCB : 6WB) had a higher pH or presented small

drops. However, cultures in smaller proportions or

without WB (6PSCB : 4WB, 8PSCB : 2WB and 10PSCB :

0WB) had drops in pH, and there were no increments to

this parameter during this experiment.

Figure 2b shows the quantity of mycelial mass estima-

ted by means of quantity of N-acetylglucosamine. It was

found that up to the fourth day of culture, all cultures

with WB in their composition indicated higher amounts

of mycelial mass than the culture that contained only

PSCB (10PSCB : 0WB). However, this culture (10PSCB :

0WB) presented later increments in the formation of

mycelial mass.

At the beginning of the process, treatments

6PSCB : 4WB, 4PSCB : 6WB and 2PSCB : 8WB presen-

ted higher mycelial mass values. This indicates that the

development of fungus P. echinulatum is favoured in

culture media formulated with the mixture of WB and

PSCB.

Discussion

Analysing the enzymatic activities found in this study, it

can be concluded that the production of FPA, endoglu-

canases, b-glucosidases and xylanases is favoured in SSFin media formulated using mixtures of PSCB and WB.

Thus, these substrates are feasible alternative sources of

enzyme production by P. echinulatum strains. It is also

found that a given enzyme of the cellulose complex can

be induced, simply alternating the proportion of sub-

strates in the culture medium.

The main FPA obtained was 3289 190 U gdm)1 bytreatment 2PSCB : 8WB on the third day. Comparing this

value to values available in the literature (Table 1), it is

seen that fungus P. echinulatum has a high potential for

FPA secretion.

The highest b-glucosidase activities were 5895 258U gdm)1, obtained on the fourth day by 2PSCB : 8WB.

The highest activity for endoglucanase was 28236 124U gdm)1, for treatment 6PSCB : 4WB on the fourth day,

while the highest xylanase activities were around

10 U gdm)1, these activities were found on the second to

fourth days of culture in several cultures.

It is not always possible to compare b-glucosidase,endoglucanase and xylanases reported in the literature,

because of lack of standard methods to determine the

activity of these enzymes. However, Table 2 shows that

some studies were performed with different substrates for

enzyme production and others for the determination of

enzymatic activity.

According to the data in this table, the b-glucosidaseactivities found for P. echinulatum are high compared

with most papers cited in Table 2. Only micro-organisms

Thermoascus aurantiacus and Penicillium decumbens were

more active. As to endoglucanase activity, once again the

potential of P. echinulatum for enzyme production is

seen, but only micro-organism T. aurantiacus presented

higher activity. However, when the activities of xylanases

obtained for P. echinulatum are compared, it is found

that they are lower than the other fungal lines. It is sug-

gested that xylanases be produced with P. echinulatum,

using sugar cane bagasse that is untreated or has under-

gone some pretreatment that will prevent loss of hemicel-

lulose. In pretreatment of the sugar cane bagasse used in

this study, after autoclaving, the substrate was washed to

remove sodium hydroxide and lignin, but at this stage

part of the hemicellulose was lost.

The xylanase activities may have been induced by the

presence of cellulose According to Olsson et al. (2003),

Trichoderma reesei presented high levels of endoxylanase

when grown in cellulase; and also according to Aro

et al. (2001), the presence of cellulose may induce not

only cellulase production but also xylanases, because

0 1 2 3 4 5 62

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Myc

elia

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m

1

(a)

(b) 10PSCB:0WB8PSCB:2WB

6PSCB:4WB

4PSCB:6WB

2PSCB:8WB

0PSCB:10WB

Figure 2 Variation of the pH (a) and mycelial mass (b) in media for-

mulated with different proportions of solid-state pretreated cane

bagasse and wheat bran, using strain 9A02S1 of Penicillium echinula-

tum. PSCB: pretreated sugar cane bagasse, WB: wheat bran. The

numbers shown in the legends indicate the proportion of each med-

ium component used.



the cellulase regulator, ACEII, also affects xylanase regu-

lation.

Moreover, according to the results, the secretion of P.

echinulatum enzymes contains all the enzymes of the cel-

lulose complex and also secretes xylanases, besides having

good thermal stability of FPA and b-glucosidase enzymesat 50C (Camassola et al. 2004). It is a valuable character-istic for its application in processes such as the enzymatic

hydrolysis of cellulose and lignocellulose to production

glucose syrup.

According to the results obtained, it is suggested that

the main activities detected in the media formulated with

WB and PSCB mixtures, besides the composition of the

inducer substrate present in the bagasse, may be due to

greater aeration of these cultures, as it is well known that

cultures in media containing only WB. The medium

packaging makes it difficult to transfer oxygen and thus

diminishes the development of the fungal mass. This

assumption is corroborated by determinations of the

mycelial mass (Fig. 2b), where treatments 6PSCB : 4WB,

4PSCB : 6WB and 2PSCB : 8WB showed higher determi-

nations of mycelial mass.

According to Poorna and Prema (2007), the size of the

WB particles influenced enzyme and biomass production.

This influence is due to the agglomeration of particles

that could inhibit oxygen transfer. Micro-organism adher-

ence and penetration, as well as the action of the

enzymes, depend on the physical properties of the sub-

strate, such as its crystalline and amorphous nature, the

accessibility area, surface, area, porosity, particle size, etc.

(Krishna 2005).

The substrate mixture makes more nutrients available

for mycelial development and the presence of inducer

substances for enzyme production. In Fomes sclero-

dermeus, the mixture of substrates (soy WB 1 : 1)induced the highest levels of hydrolases, the differences

were more evident in polygalacturonase and polymethyl-

galacturonase activities (Papinutti and Forchiassin 2007).

This has a result is similar to that obtained with T. reesei

where the resulting enzyme activities were generally

higher during the growth on mixed substrates than those

obtained when only a single substrate was used (Olsson

et al. 2003).

According to Archana and Sathyanarayana (1997), WB

is universally suitable as substrate because it contains suf-

ficient nutrients and remains free even under high mois-

ture conditions, providing a large surface. The

biochemical composition of WB indicated that when this

material was hydrolysed, it contained a considerable

amount of soluble sugar like glucose (425% dry wt),xylose (154% dry wt), arabinose (31% dry wt) andgalactose (27% dry wt), required to initiate micro-organ-ism growth and replication. The degree of substitution of

the main xylan chains by arabinose was higher in WB

(Lequart et al. 1999). It contained hemicellulose (45%),

which may act as an inducer, and organic nitrogen

sources (23%) that are essential for protein synthesis

(Babu and Satyanarayana 1996).

Still, the use of lignocellulosic materials such as PSCB

for enzyme production has advantages compared with

the SmF: high production of enzymes using a low-cost

media (Viniegra-Gonzales et al. 2003) and the possible

use of the bioconverted substrate because of its

increased digestibility (Mukherjee and Nandi 2004).

When the substrate is based on a high protein content

raw material such as WB, its nutritional value is

increased and this material can be potentially used in

animal feed.

Table 1 Comparisons of FPA production

from different fungi grown on lignocellulosic

materials

Fungi Substrate FPA (U g)1) References

P. echinulatum 9A02S1 Wheat bran and pretreated

sugar cane bagasse

3289 This work

Trichoderma harzianum Wheat straw and bran 18 Deschamps et al. (1985)

Penicillium decumbens Wheat straw and

wheat bran

177 Mo et al. (2004)

T. reesei LM-UC 4 and

Aspergillus phoenicis

Sugar cane bagasse 134 Gutierrez-Correa and

Tengerdy (1997)

T. reesei LM-UC 4E1 Sugar cane bagasse 10 Gutierrez-Correa and

Tengerdy (1997)

Thermoascus aurantiacus Dry wheat straw 55 Kalogeris et al. (2003)

T. reesei LM-UC 4 Sugar cane bagasse 53 Gutierrez-Correa and

Tengerdy (1997)

Myceliophthora sp. Rice straw 244 Badhan et al. (2007)

Myceliophthora sp. Wheat straw 137 Badhan et al. (2007)

Myceliophthora sp. Wheat bran 074 Badhan et al. (2007)

Myceliophthora sp. Bagasse 07 Badhan et al. (2007)

Myceliophthora sp. Corn cob 031 Badhan et al. (2007)



Table 2 Comparisons of b-glucosidase, endoglucanase and xylanase productions from different fungi grown on lignocellulosic materials

Fungi Substrate b-glucosidase (U g)1) References


sugar cane bagasse

5895 This work

T. aurantiacus Wheat straw 79 Kalogeris et al. (2003)

P. decumbens Wheat straw and wheat bran 528 Mo et al. (2004)

T. reesei and A. phoenicis Sugar cane bagasse 181 Gutierrez-Correa and

Tengerdy (1997)

T. reesei Sugar cane bagasse 91 Gutierrez-Correa and

Tengerdy (1997)


Tengerdy (1997)






Fungi Substrate Endoglucanase (U g)1) References


sugar cane bagasse

28236 This work


T. harzianum Wheat straw and bran 198 Deschamps et al. 1985

T. reesei and A. phoenicis Sugar cane bagasse 738 Gutierrez-Correa and

Tengerdy (1997)


Tengerdy (1997)

Sporotrichum pulverulentum Rice straw 20 Shamala and Sreekantiah (1986)


Tengerdy (1997)

A. niger Wheat straw and wheat bran 148 Jecu (2000)

A. ustus Rice straw and wheat bran 14 Shamala and Sreekantiah (1986)






Fungi Substrate Xylanase (U g)1) References


sugar cane bagasse

10 This work

T. lanuginosus D2 W3 Sorghum straw 48 000 Sonia et al. (2005)


P. themophila J18 Wheat straw 7745 Yang et al. (2006)


T. aurantiacus Bagasse 2700 Souza et al. (1999)

T. aurantiacus Sugar cane bagasse 1597 Milagres et al. (2004)




Myceliophthora sp Corn cob 4116 Badhan et al. (2007)

Trichoderma viride TS Sugar beet pulp 200 Grajek and Gervais (1987)


Aspergillus awamori Grape pomace 38 Botella et al. (2007)



Conclusion

The present work established the potential of P. echinula-

tum for FPA, endoglucanase, b-glucosidase and xylanaseproduction in SSF, indicating that WB may be partially

substituted by the PSCB. The incorporation of cheap

sources, such as sugar cane bagasse, into media for the

production of lignocellulose enzymes should contribute to

a decrease in the costs of the production of enzymatic

complexes capable of hydrolysing lignocellulose residues

for the formation of fermented syrups, thus contributing

to the economic production of bioethanol.

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